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Expansion of Human Airway Basal Stem Cells and Their Differentiation as 3D Tracheospheres

Hynds, RE; Butler, CR; Janes, SM; Giangreco, A; (2016) Expansion of Human Airway Basal Stem Cells and Their Differentiation as 3D Tracheospheres. Methods in Molecular Biology pp. 1-11. 10.1007/7651_2016_5. (In press). Green open access

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Abstract

Although basal cells function as human airway epithelial stem cells, analysis of these cells is limited by in vitro culture techniques that permit only minimal cell growth and differentiation. Here, we report a protocol that dramatically increases the long-term expansion of primary human airway basal cells while maintaining their genomic stability using 3T3-J2 fibroblast coculture and ROCK inhibition. We also describe techniques for the differentiation and imaging of these expanded airway stem cells as three-dimensional tracheospheres containing basal, ciliated, and mucosecretory cells. These procedures allow investigation of the airway epithelium under more physiologically relevant conditions than those found in undifferentiated monolayer cultures. Together these methods represent a novel platform for improved airway stem cell growth and differentiation that is compatible with high-throughput, high-content translational lung research as well as human airway tissue engineering and clinical cellular therapy.

Type: Article
Title: Expansion of Human Airway Basal Stem Cells and Their Differentiation as 3D Tracheospheres
Location: United States
Open access status: An open access version is available from UCL Discovery
DOI: 10.1007/7651_2016_5
Publisher version: http://dx.doi.org/10.1007/7651_2016_5
Language: English
Additional information: Copyright © 2016 Springer Science+Business Media New York. The final publication is available at Springer via http://dx.doi.org/10.1007/7651_2016_5.
Keywords: Adult stem cells, Cell culture techniques, Cilia, Epithelial cells, Goblet cells, Lung, Primary cell culture, Stem cells
UCL classification: UCL > Provost and Vice Provost Offices
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Cancer Institute
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Cancer Institute > Research Department of Oncology
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Div of Medicine
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Div of Medicine > Respiratory Medicine
URI: https://discovery.ucl.ac.uk/id/eprint/1508754
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